LncRNA KCNQ1OT1通过调控 miR-506-3p表达调节喉鳞状细胞癌细胞的生物学行为

目的 探讨长链非编码RNA(LncRNA)KCNQ1重叠转录物1(KCNQ1OT1)对喉鳞状细胞癌细胞生物学的影响及作用机制.方法 于2019年5月至2020年2月,采用RT-qPCR法检测了45例来自山东大学齐鲁医院桓台分院喉鳞状细胞癌病人的癌组织和癌旁组织及购自上海研生实业有限公司的人胚肺成纤维细胞WI-38和购自中国科学院上海细胞库的喉鳞状细胞癌细胞系(EV-SCC-18、AMC-HN-8和HCC345)中KCNQ1OT1和miR-506-3p表达.以HCC345细胞为研究对象,转染KCNQ1OT1小干扰RNA或共转染KCNQ1OT1小干扰RNA与miR-506-3p抑制剂至HCC345细胞后,MTT法、流式细胞术、Transwell分别检测细胞增殖、凋亡、迁移和侵袭.双荧光素酶报告基因实验验证KCNQ1OT1与miR-506-3p的调控关系.结果 喉鳞状细胞癌组织中KCNQ1OT1表达高于癌旁组织(0.81±0.09)比(0.27±0.08)],miR-506-3p表达低于癌旁组织(0.24±0.08)比(0.83±0.09)].喉鳞状细胞癌细胞系(EV-SCC-18、AMC-HN-8和HCC345)中KCNQ1OT1表达均高于WI-38细胞(2.44±0.22)、(2.69±0.21)、(3.61±0.24)比(1.00±0.13)],miR-506-3p表达均低于WI-38细胞(0.41±0.13)、(0.30±0.12)、(0.22±0.11)比(1.00±0.12)].与未敲减KCNQ1OT1的HCC345细胞比较,敲减KCNQ1OT1的HCC345细胞活力(0.41±0.06)比(0.81±0.12)]、迁移数(86.32±15.21)个比(162.31±20.23)个]和侵袭数(65.23±12.05)个比(140.26±18.27)个]均降低,细胞凋亡率升高(34.13±3.60)％比(3.79±2.37)％].KCNQ1OT1在HCC345细胞中负调控miR-506-3p表达.敲减miR-506-3p逆转敲减KCNQ1OT1对HCC345细胞增殖、凋亡、迁移和侵袭的影响.结论 KCNQ1OT1在喉鳞状细胞癌组织和细胞系中表达升高,其通过靶向miR-506-3p促进喉鳞状细胞癌细胞的恶性生物学行为.

LncRNA KCNQ1OT1 regulates biological behavior of laryngeal squamous cell carcinoma cells by regulating miR-506-3p expression

Objective To investigate the effect and mechanism of LncRNA KCNQ1OT1 on the biological behavior of laryngeal squa-mous cell carcinoma cells.Methods From May 2019 to February 2020, the expression levels of KCNQ1OT1 and miR-506-3p in 45 cas-es of cancer tissues and para-cancerous tissues from patients with laryngeal squamous cell carcinoma from the Huantai Branch of Shan-dong University Qilu Hospital, and human embryonic lung fibroblasts WI-38 purchased from Shanghai Yansheng Industrial Co., Ltd. and laryngeal squamous cell carcinoma cell lines (EV-SCC-18, AMC-HN-8 and HCC345) purchased from Shanghai Cell Bank of the Chinese Academy of Sciences were detected by RT-qPCR. HCC345 cells were used as the research object and transfected with KCNQ1OT1 small interfering RNA or co-transfected with KCNQ1OT1 small interfering RNA and miR-506-3p inhibitor. MTT, flow cytometry and Transwellwere used to detect the proliferation, apoptosis, migration and invasion. The double luciferase reporter gene experiment verified the regu-latory relationship between KCNQ1OT1 and miR-506-3p.Results The expression of KCNQ1OT1 in laryngeal squamous cell carcinoma was higher than that in adjacent tissues (0.81±0.09) vs. (0.27±0.08)], but the expression of miR-506-3p was decreased lower than that in adjacent tissues (0.24±0.08) vs. (0.83±0.09)]. The expression of KCNQ1OT1 in laryngeal squamous cell carcinoma cell lines (EV-SCC-18, AMC-HN-8 and HCC345) were higher than that in WI-38 cells (2.44±0.22), (2.69±0.21), (3.61±0.24) vs. (1.00±0.13)], but the expres-sion of miR-506-3p were lower than that of WI-38 cells (0.41±0.13), (0.30±0.12), (0.22±0.11) vs. (1.00±0.12)]. Compared with HCC345 cells without knocking down KCNQ1OT1, the viability (0.41±0.06) vs. (0.81±0.12)], the number of migration (86.32±15.21) vs. (162.31± 20.23)], and the number of invasion (65.23±12.05) vs. (140.26±18.27)] of HCC345 cells that knocked down KCNQ1OT1 were reduced, but cell apoptosis was increased (34.13±3.60)% vs. (3.79±2.37)%]. KCNQ1OT1 negatively regulated miR-506-3p in HCC345 cells. Knockdown of miR-506-3p reversed the effect of knocking down KCNQ1OT1 on the proliferation, apoptosis, migration and invasion of HCC345 cells.Conclusion KCNQ1OT1 is highly expressed in laryngeal squamous cell carcinoma tissues and cell lines. It promotes themalignant biological behavior of laryngeal squamous cell carcinoma cells by targeting the expression of miR-506-3p.